Most sudden cardiac deaths occur in patients with coronary artery disease and associated left ventricular dysfunction. Epicardial coronary artery abnormalities resulting in acute or chronic ischemic insults account for up to 80% of clinical arrhythmias. Randomized trials and clinical electrophysiological studies have demonstrated the ineffectiveness of anti-arrhythmic drug therapy in reducing mortality in this high-risk patient population. Paradoxically, conventional pharmacotherapies targeting ion channel proteins are often associated with increased rather than decreased mortality, possibly due to a potent pro-arrhythmic effect of these drugs. Numerous studies have established the importance of abnormal intracellular calcium (Ca2+) cycling in mechano-electrical dysfunction. Defective sequestration of Ca2+ by the sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a), coupled with increased diastolic SR Ca2+ leak via the ryanodine receptor (RYR2), result in cytosolic Ca2+ overload and associated dysfunction in ischemic heart disease. Key molecular targets that modulate SR Ca2+uptake and release include: 1) SERCA2a and its newly discovered post-translational modification by SUMO1, 2) Phospholamban (PLB), an endogenous inhibitor of SERCA2a, 3) FKBP12.6, a key component of the RYR2 macromolecular complex which stabilizes RYR2 activity, and 4) CAMKIIc, a serine/threonine protein kinase which regulates intracellular Ca2+ cycling, including SR Ca2+ leak through RYR2 phosphorylation. The development of novel gene-based therapies that target these central components of intracellular Ca2+ cycling requires the investigation of their electrophysiological consequences, including pro- arrhythmic risk, in clinically relevant large animal models that closely mimic human ischemic heart disease. A major obstacle that has hindered the translation of these potentially effective molecular therapies has been the availability of adequate vectors for long-term gene transfer. Although AAV vectors were found to be safe in multiple clinical trials, their widespread use for gene delivery is limited by: 1) non-specificity to the heart and 2) pre-existing neutralizig antibodies to conventional AAV serotypes in <50% of candidates. A major innovation of the current application is the proposed use of chimeric AAV based bionanoparticles that exhibit superior cardiac tropism while escaping inherent immunological limitations in patients. We will take advantage of clinically relevant porcine models and gene delivery systems to test the central hypothesis that: a) SUMO1 SERCA2a overexpression, b) PLB silencing, c) FKBP12.6 overexpression, and d) CAMKIIc inhibition are associated with distinct electrophysiological consequences in preclinical models of CAD. These studies will reveal the electrophysiological benefits and potential pitfalls associated with novel (e.g. SERCA2a + SUMO1) molecular therapies for CAD.